To establish and optimize the genetic transformation technology of Penicillium marneffei by Agrobacterium tumefaciens-mediated transformation
LI Xinlei
LI Xin
CAO Cunwei
Abstract：Objective To construct and optimize the condition of Agrobacteriumtumefaciens?mediated transformattechology in Penicillium marneffei.Method A.tumefaciens transformed by Calcium chloride with pDHt/ SK::pyrG plasmids insert pyrG gene into SPM4 (pyrG?,niaD?),medium without uracil screening positive transformat,Using PCR verification restructuring.It aimed at finding out the optimal protocol by changing some important effecting factors including A.tumefaciens strains,concentration,transformat medi?um,co?cultivation temperature,period,AS. The positive transformant growed well in the media without uracil, and PCR analysis showed that T?DNA was inserted into the transformants. Result The transformation efficiency was about 300 transformants/ 106 spores by optimizing.The optimal condition for AGL?1,concentration of A.tumefaciens OD600 = 0.8,the AS concentration of 200 μmol/L,without nitrocellulose filters,co?culture 48 h in 25℃ .Conclusion A.tumefaciens?mediated transformattechology in P.marneffei.was successfully established,simplified and optimized.The method can be used for PM gene function research.
Keywords：Penicillium marneffeiAgrobacterium tumefacienstransformation
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 88-91 )